Analysis of GPCR localization and trafficking

Methods Mol Biol. 2011;746:425-40. doi: 10.1007/978-1-61779-126-0_25.

Abstract

Localization and trafficking of G protein-coupled receptors (GPCRs) is increasingly recognized to play a fundamental role in receptor-mediated signaling and its regulation. Individual receptors, including closely homologous subtypes with otherwise similar functional properties, can differ considerably in their membrane trafficking properties. In this chapter, we describe several approaches for experimentally assessing the subcellular localization and trafficking of selected GPCRs. Firstly, we describe a flexible method for receptor localization using fluorescence microscopy. We then describe two complementary approaches, using fluorescence flow cytometry and surface biotinylation, for examining receptor internalization and trafficking in the endocytic pathway.

MeSH terms

  • Biotinylation
  • Flow Cytometry
  • HEK293 Cells
  • Humans
  • Intracellular Space / metabolism*
  • Protein Transport
  • Receptors, G-Protein-Coupled / metabolism*
  • Staining and Labeling
  • Tissue Fixation

Substances

  • Receptors, G-Protein-Coupled